An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE)
The non-stereospecific α-haloalkanoic acid dehalogenase E (DehE) degrades many halogenated compounds but is ineffective against β-halogenated compounds such as 3-chloropropionic acid (3CP). Using molecular dynamics (MD) simulations and site-directed mutagenesis we show here that introducing the muta...
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author | Abdul Hamid, Azzmer Azzar Tengku Abdul Hamid, Tengku Haziyamin Abdul Wahab, Roswanira Omar, Mohd. Shahir Shamsir Huyop, Fahrul |
author_facet | Abdul Hamid, Azzmer Azzar Tengku Abdul Hamid, Tengku Haziyamin Abdul Wahab, Roswanira Omar, Mohd. Shahir Shamsir Huyop, Fahrul |
author_sort | Abdul Hamid, Azzmer Azzar |
collection | ePrints |
description | The non-stereospecific α-haloalkanoic acid dehalogenase E (DehE) degrades many halogenated compounds but is ineffective against β-halogenated compounds such as 3-chloropropionic acid (3CP). Using molecular dynamics (MD) simulations and site-directed mutagenesis we show here that introducing the mutation S188V into DehE improves substrate specificity towards 3CP. MD simulations showed that residues W34, F37, and S188 of DehE were crucial for substrate binding. DehE showed strong binding ability for D-2-chloropropionic acid (D-2CP) and L-2-chloropropionic acid (L-2CP) but less affinity for 3CP. This reduced affinity was attributed to weak hydrogen bonding between 3CP and residue S188, as the carboxylate of 3CP forms rapidly interconverting hydrogen bonds with the backbone amide and side chain hydroxyl group of S188. By replacing S188 with a valine residue, we reduced the inter-molecular distance and stabilised bonding of the carboxylate of 3CP to hydrogens of the substrate-binding residues. Therefore, the S188V can act on 3CP, although its affinity is less strong than for D-2CP and L-2CP as assessed by Km. This successful alteration of DehE substrate specificity may promote the application of protein engineering strategies to other dehalogenases, thereby generating valuable tools for future bioremediation technologies. |
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institution | Universiti Teknologi Malaysia - ePrints |
language | English |
last_indexed | 2024-03-05T19:40:30Z |
publishDate | 2015 |
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spelling | utm.eprints-577942021-08-04T05:07:17Z http://eprints.utm.my/57794/ An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE) Abdul Hamid, Azzmer Azzar Tengku Abdul Hamid, Tengku Haziyamin Abdul Wahab, Roswanira Omar, Mohd. Shahir Shamsir Huyop, Fahrul QD Chemistry The non-stereospecific α-haloalkanoic acid dehalogenase E (DehE) degrades many halogenated compounds but is ineffective against β-halogenated compounds such as 3-chloropropionic acid (3CP). Using molecular dynamics (MD) simulations and site-directed mutagenesis we show here that introducing the mutation S188V into DehE improves substrate specificity towards 3CP. MD simulations showed that residues W34, F37, and S188 of DehE were crucial for substrate binding. DehE showed strong binding ability for D-2-chloropropionic acid (D-2CP) and L-2-chloropropionic acid (L-2CP) but less affinity for 3CP. This reduced affinity was attributed to weak hydrogen bonding between 3CP and residue S188, as the carboxylate of 3CP forms rapidly interconverting hydrogen bonds with the backbone amide and side chain hydroxyl group of S188. By replacing S188 with a valine residue, we reduced the inter-molecular distance and stabilised bonding of the carboxylate of 3CP to hydrogens of the substrate-binding residues. Therefore, the S188V can act on 3CP, although its affinity is less strong than for D-2CP and L-2CP as assessed by Km. This successful alteration of DehE substrate specificity may promote the application of protein engineering strategies to other dehalogenases, thereby generating valuable tools for future bioremediation technologies. Penerbit UTM Press 2015 Article PeerReviewed application/pdf en http://eprints.utm.my/57794/1/RoswaniraAbdulWahab2015_AnS188VMutationAltersSubstrateSpecificity.pdf Abdul Hamid, Azzmer Azzar and Tengku Abdul Hamid, Tengku Haziyamin and Abdul Wahab, Roswanira and Omar, Mohd. Shahir Shamsir and Huyop, Fahrul (2015) An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE). Plos One, 10 (3). pp. 1-21. ISSN 1932-6203 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0121687 |
spellingShingle | QD Chemistry Abdul Hamid, Azzmer Azzar Tengku Abdul Hamid, Tengku Haziyamin Abdul Wahab, Roswanira Omar, Mohd. Shahir Shamsir Huyop, Fahrul An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE) |
title | An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE) |
title_full | An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE) |
title_fullStr | An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE) |
title_full_unstemmed | An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE) |
title_short | An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE) |
title_sort | s188v mutation alters substrate specificity of non stereospecific alpha haloalkanoic acid dehalogenase e dehe |
topic | QD Chemistry |
url | http://eprints.utm.my/57794/1/RoswaniraAbdulWahab2015_AnS188VMutationAltersSubstrateSpecificity.pdf |
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